Mean Number Of Somatic Embryos Research Articles

Induction of direct somatic embryogenesis and shoot organogenesis and histological study in tree peony (Paeonia sect. Moutan)

Tree peony (Paeonia sect. Moutan) is a world famous ornamental and economically important species, but it is recalcitrant to in vitro regeneration. Here, we present a protocol for induction of direct somatic embryogenesis (SE) and direct shoot organogenesis (SO) from zygotic embryos (ZEs) of Paeonia rockii and P. ostii. The results showed that explant genotypes, ZEs stages, sucrose and plant growth regulators (PGRs) have greatly influences on in vitro morphogenesis. The highest frequency of SE (48%) and mean number of somatic embryos (5) was obtained from mature ZE of P. rockii ‘Jing Hong’ when cultured on modified Murashige and Skoog (mMS) medium supplemented with 2.22 µM BA and 0.23 M sucrose.The proliferation can be achieved by recurrent production of embryogenic callus and somatic embryos on mMS medium supplemented with 0.89 µM BA. The germinated rate of somatic embryos was 45% on WPM medium containing 2.22 µM BA and 1.44 µM GA3 after dormancy release at 4 °C under dark for 20 days. In addition, direct SO was obtained firstly in tree peony by using a variety of cytokinins. Morpho-histological analysis confirmed the initiation, development and reserve accumulation of somatic embryos and shoots. The study will be benificial to the propagation and breeding of tree peony. Tree peony (Paeonia sect. Moutan) is a world famous ornamental and economically important species which is recalcitrant to in vitro regeneration. Direct somatic embryogenesis protocol of P. rockii ‘Jing Hong’ was developed and morpho-histological analysis confirmed the initiation, development and reserve accumulation during in vitro morphogenesis. The study will provide valuable reference for propagation and breeding of tree peony.

Induction of somatic embryogenesis and evaluation of genetic stability in regenerated plants of Magnolia dealbata

The utility of plant tissue culture for the mass propagation of trees is well known, but continuous in vitro multiplication of plant material may increase the possibility of somaclonal variation; therefore, it is essential to evaluate the genetic integrity of regenerants from species-specific in vitro protocols prior to mass production and implementation. The objectives of this study were: 1) to determine the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) concentration over two cycles of secondary somatic embryogenesis in Magnolia dealbata; and 2) to verify the genetic stability of the regenerants obtained. The embryogenic response was not significantly affected by the concentration of 2,4-D but did vary across cycles of induction. The addition of 4.52 μM 2,4-D induced the highest total number of embryos (100.5), the mean number of somatic embryos (25.1) and somatic embryos per explant (80.6). In both 2,4-D concentration (2.26 or 4.52 μM), genetic integrity between the donor and the propagated clones was 0.90, and the low genetic instability (≤ 0.10 in both PGR treatments) might be due to effect of cyclic somatic embryogenesis or the different response of the explants at stress in in vitro culture conditions. However, it is necessary to examine more cell lines and somatic embryogenesis cycles.

Recurrent somatic embryogenesis and development of somatic embryos in Akebia trifoliata (Thunb.) Koidz (Lardizabalaceae)

A simple and effective protocol was established for recurrent somatic embryogenesis and plant regeneration in Akebia trifoliata (Thunb.) Koidz. Somatic embryos directly formed from the root zone of the immature zygotic embryos cultured on MS medium devoid of plant growth regulator (PGR). The induction frequency of immature zygotic embryos was 52.5%, and the mean number of somatic embryos reached up to 19.5. Secondary somatic embryos arose on the primary embryos at a high frequency (95.8%), and this process maintained in a recurrent way on PGR-free MS medium. The highest mean number of somatic embryos also appeared in the primary embryos which can reach up to 44.7. While both of the embryogenic potential and mean number of embryos per explant displayed a gradual diminution with subculturing. The addition of 0.5 mg l−1 6-benzyladenine in the 1/2 MS regeneration medium can significantly improve the frequency of somatic embryos convert to plantlets. The microscopic analysis revealed that the development process of somatic embryos via the globular, heart, torpedo and cotyledonal stages in A. trifoliata, the histological analysis showed that somatic embryos directly initiated from root epidermal cells and there are no vascular connections between the somatic embryos and maternal tissue. Regenerated plantlets acclimated successfully to greenhouse conditions. Approximately 65% plantlets survived and displayed no morphological characteristics differences with seed-derived plants. The simple and effective protocols established in this study will promote large-scale clonal propagation and genetic improvement of A. trifoliata. Somatic embryogenesis has never been reported in Akebia trifoliata (Thunb.) Koidz. The study showed the origin and development process of somatic embryos in Akebia trifoliata (Thunb.) Koidz.

Somatic embryogenesis and plant regeneration of cassava (Manihot esculenta Crantz) landraces from Cameroon.

A procedure to regenerate cassava (Manihot esculenta Crantz) cultivars from Cameroon via somatic embryogenesis (SE) was developed. Shoot apical meristems and immature leaf lobes were used as explants on Murashige and Skoog (MS) basal medium containing 33 or 50 µM of the auxins Picloram (Pic), 2,4-Dichlorophenoxyacetic acid (2,4-D), Dicamba (Dic), and α-Naphthalene acetic acid. Cultivar performance was assessed using SE and number of somatic embryos produced. Overall, the frequency of primary somatic embryogenesis (PSE) and the mean number of somatic embryos produced varied considerably with genotype, type of auxin and concentration tested. For example, cultivar (cv.) Ngan Mbada showed the best performance on MS medium supplemented with 50 µM Pic with a SE frequency of 40 % and an average number of somatic embryos of 90. The second best performance was recorded in cv. Local Red on MS medium supplemented with 33 µM 2,4-D, where the SE frequency was 40 % and an average number of somatic embryos of 60.5. Cultivar Ekona Red recorded the best performance on medium supplemented with 50 µM Pic showing a SE frequency of 47 % and an average number of somatic embryos of 45. We further examined secondary and cyclic somatic embryogenesis (SSE, CSE) and both were also observed to vary with genotype, however, both exhibited significantly higher frequencies of SE compared with PSE. SE started to decline at the fourth cycle of embryogenesis. Examination of organogenesis showed that shoot bud induction from green cotyledons varied across cultivars and benzylaminopurine was shown to outperform Thidiazuron in the ability to induce organogenesis. Furthermore, the frequencies of bud induction were identical under light and dark conditions. Finally, regenerated plants grew easily in the greenhouse with 90–100 % survival rate and did not display detectable variation in morphology.

Embryogenesis, plant regeneration and cardiac glycoside determination in Digitalis ferruginea subsp. ferruginea L

The present study reports, for the first time, an efficient in vitro plant regeneration protocol for Digitalis ferruginea subsp. ferruginea L. (rusty foxglove). We have used different concentrations of gibberellic acid (GA3) on Murashige and Skoog (MS) medium to assess the germination frequency of seeds. High frequency of germination was achieved on MS medium with 1.0 mg l−1 GA3. 6-Benzylaminopurine (BAP) combined with α-naphtaleneacetic acid (NAA) or 2, 4-dichlorophenoxy acetic acid (2, 4-D) in the induction MS medium induced both somatic embryogensis and shoot organogenesis. The highest percentage of callus growth (85 %) was obtained when hypocotyl explants were cultured on MS medium containing 0.5 mg l−1 2, 4-D plus 1.0 mg l−1 BAP. The maximum mean number of somatic embryos (7.3 ± 1.3 embryos) or shoots (12.0 ± 1.1 shoots) per callus was obtained when medium contained 0.25 mg l−1 NAA plus 1.0 mg l−1 BAP or 0.5 mg l−1 NAA plus 2.0 mg l−1 BAP. The regenerated shoots easily rooted on MS medium. Higher amounts of lanatoside C [13.2 ± 0.5 mg 100 g−1 dry weight (dw)] and digoxin (2.93 ± 0.31 mg 100 g−1 dw) accumulation were obtained when shoots were obtained by indirect regeneration. We also investigated derivatives of cardenolides, i.e., digitoxigenin (730 ± 180 mg 100 g−1 dw), gitoxigenin (50 ± 20 mg 100 g−1 dw) and digoxigenin (490 ± 170 mg 100 g−1 dw) from natural samples.

Plant regeneration through direct somatic embryogenesis from immature zygotic embryos of the medicinal plant, Paris polyphylla Sm.

A protocol for induction of direct somatic embryogenesis and subsequent plant regeneration for the medicinally important and endangered plant Paris polyphylla Sm. has been developed for the first time. Immature zygotic embryos (IZEs) were cultured on different media namely Gamborg (B5), ½ B5, Murashige and Skoog (MS), ½ MS, Chu et al. (N6), ½ N6, Schenk and Hildebrandt (SH) and ½ SH. Highest frequency of somatic embryogenesis (32.6 %) and mean number of somatic embryos (SEs) per explant (28.7 ± 1.7) were obtained on ½ MS medium directly without an intermediate callus phase. The frequency of SE induction was significantly increased to 40.7 % when ½ MS medium was solidified with gelrite compared to agar (32.6 %). Secondary somatic embryos (SSEs) appeared on the primary SEs in a repetitive way on plant growth regulator-free ½ MS medium but with a gradual decrease in embryogenic potential during subsequent subcultures. Plasmolyzing pre-treatment of SSEs with 1.0 M mannitol for 12 h effectively maintains its embryogenic capacity. Primary embryos at the elongated dimpled and early cotyledonary stage displayed the highest embryo forming capacity of 26.94 and 27.87, respectively. High frequency of SE germination (94.0 %) occurred on ½ MS medium with 0.5 mg/l gibberellic acid. Highest percentage of seedling to plantlet conversion was observed in the medium supplemented with 0.05 mg/l 6-benzylaminopurine and 0.1 mg/l α-naphthalene acetic acid. Regenerated plants displayed morphological characteristics similar to that of the wild plants. Flow cytometry analysis showed ploidy stability of the regenerated plants.

Somatic embryogenesis in Alnus glutinosa (L.) Gaertn

Induction of somatic embryos and plant regeneration was demonstrated for the first time in Alnus glutinosa. Somatic embryos were initiated from zygotic embryos collected 1–3 weeks post-anthesis (WPA), i.e., when they were at globular or early cotyledonary stage and were 0.5–1 mm in length. Induction frequency (16.6 %) and the mean number of somatic embryos (4.5 embryos/explant) were highest after culture of zygotic embryos, collected at 3 WPA, on Murashige and Skoog medium (MS) supplemented with 0.9-μM 2,4-dichlorophenoxyacetic acid and 2.22-μM benzyladenine (BA). No embryogenic induction was observed on medium with BA alone. Initial somatic embryos differentiated indirectly from callus tissue formed at the surface of the zygotic embryos. Embryogenic competence was maintained by secondary embryogenesis, which was affected by explant type, plant growth regulators and genotype. Secondary embryogenesis was induced by culture of small groups of whole somatic embryos or isolated cotyledon explants on medium consisting of MS medium (half-strength macronutrients) supplemented with 0.44-μM BA. Histological study of isolated cotyledon explants revealed that secondary embryos developed directly from differentiated embryogenic tissue on the surface of cotyledons. Somatic embryos at successive stages of development, including cotyledonary-stage embryos with shoot and root meristems, were evident. For plantlet conversion, somatic embryos were transferred to maturation medium supplemented with 3 % maltose, followed by 6 weeks of culture in Woody Plant Medium supplemented with 0.44-μM BA and 0.46-μM Zeatin (Z). This novel protocol appears promising for mass propagation, conservation and genetic transformation of black alder.

Hygromycin promotes somatic embryogenesis in spinach

Hygromycin (hyg) at low doses (0.5–1.0 mg l−1) promoted somatic embryogenesis from apical sections of spinach lateral roots. The highest promoting effect on both the frequency of regeneration and the mean number of somatic embryos (SE) per explant was achieved at 0.5 mg l−1 hyg. With increasing the concentration of hyg to 1 mg l−1, the regeneration frequency decreased, while the mean SE number remained significantly higher than in control (hyg-free medium). Complete inhibition of SE regeneration started at 7.5 mg l−1 hyg. Moreover, hyg efficiently promoted the process of secondary somatic embryogenesis. Compared to control, a 2.75-fold increase in the secondary somatic embryo (SSE) mean number was obtained at 0.5 mg l−1 hyg, and the increment was still discernible at 1.0 and 2.5 mg l−1 hyg. Both primary SE and SSE explants became completely necrotic at 12.5 mg l−1 hyg. Since attempts with direct selection at 20 mg l−1 hyg proved unsuccessful, the results obtained in this study suggest that a stepwise selection procedure is suitable, starting with selection at 0.5 mg l−1 hyg, to exploit the promoting effect of low hyg doses on SE regeneration from transformed cells, then gradually increasing the hyg concentration to 20 mg l−1 for final selection. Complete SE and SSE explant mortality at hyg above 12.5 mg l−1 guarantees a low possibility of escape during the selection process. This study will be useful for increasing the efficiency of transgenic plant regeneration following genetic transformation in spinach.

Effects of different ectomycorrhizal fungi on somatic embryogenesis of Abies cephalonica Loud

The effects of ectomycorrhizal (ECM) fungi, including Laccariabicolor (Maire) Orton, Laccaria laccata (Scop., Fr.) Berk. and Br., along with two strains of Pisolithus tinctorius (Pers.) Coker and Couch, on the proliferation and subsequent maturation of two embryogenic cell lines of Abies cephalonica Loud., designated lines 6 and 8, were investigated. In the presence of these ECM fungi, the proliferation of both embryogenic cell lines was inhibited. L. bicolor and P. tinctorius strain 2 resulted in the highest inhibition rates. On the other hand, cultivation of embryogenic cultures along with ECM fungi, termed a dual culture, increased radial growth of both P. tinctorius strains; whereas, L. bicolor and L. laccata did not grow as well in the presence of embryogenic cell masses. The dual culture during the proliferation period of embryogenic cells, however, enhanced the subsequent embryo formation and maturation of A. cephalonica; i.e. the capability of embryogenic cell lines to form somatic embryos as well as increasing the mean number of somatic embryos per 1 g fresh weight of embryogenic cell mass. However, levels of responses were highly dependent on the interaction between the specific embryogenic cell line and fungal strain.

Somatic embryogenesis and plant regeneration from root sections of Allium schoenoprasum L.

A protocol has been developed for somatic embryogenesis and subsequent plant regeneration in Allium schoenoprasum L. Calli were induced from root sections isolated from axenic seedlings and cultivated on media containing either Murashige and Skoog’s (MS) or Dunstan and Short’s mineral solution supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 6-benzylaminopurine (BA), 6-furfurylaminopurine (Kin) or thidiazuron (TDZ) at 1, 5 or 10 μM. The highest frequencies of callus induction were achieved on media with 5 μM 2,4-D in combination with 5 μM TDZ or 10 μM BA (78.9% and 78.4%, respectively). Calli were then transferred to 1 μM 2,4-D, where compact yellow callus turned to segmented yellowish callus with transparent globular somatic embryos at the surface. Calli that were previously grown on media with 5 μM 2,4-D in combination with 10 μM BA or 10 μM TDZ showed the highest frequencies of embryogenic callus formation (45% and 42%) as well as mean number of somatic embryos per regenerating callus. The choice of mineral solution formulation did not significantly affect callus induction or embryogenic callus formation. The embryos could complete development into whole plants on plant growth regulator (PGR)-free medium, but inclusion of Kin (0.5, 2.5 and 5 μM) in this phase improved somatic embryo development and multiplication. Subsequently transferred to 1/2 MS PGR-free medium, all embryos rooted and the survival rate of the plants in a greenhouse was 96%.

Effects of mutagens on somatic embryogenesis and plant regeneration in groundnut

The embryogenic calli (EC) were obtained from hypocotyl explants of groundnut (Arachis hypogaea L.) cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 0.5 mg dm−3 6-benzylaminopurine (BAP). The EC were exposed to γ-radiation (10–50 Gy) or treated with 1–5 mM of ethyl methane sulphonate (EMS) or sodium azide (SA). The mutated EC were subcultured on embryo induction medium containing 20 mg dm−3 2,4-D. Somatic embryos (SE) developed from these calli were transferred to MS medium supplemented with BAP (2.0 mg dm−3) and 0.5 mg dm−3 2,4-D for maturation. The well-developed embryos were cultured on germination medium consisting of MS salts with 2.0 mg dm−3 BAP and 0.25 mg dm−3 naphthaleneacetic acid (NAA). Well-developed plantlets were transferred for hardening and hardened plants produced normal flowers and set viable seeds. The fresh mass of the EC, mean number of SE per explant and regeneration percentage were higher at lower concentrations of mutagens (up to 30 Gy/3 mM). Some abnormalities in regenerated plants were observed, especially variations in leaf shape.

Rapid In vitro Multiplication of Chirita longgangensis W.T. Wang: An Endemic and Endangered Gesneriaceae Species in China

Experiments were conducted to establish an efficient protocol for micropropagation of Chirita longgangensis W.T. Wang. Somatic embryos formed directly at the cut edges of C. longgangensis leaf explants on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BA) and α-naphthalene acetic acid (NAA). During the somatic embryo induction stage, leaf explants and basal leaf explants were used. Leaves were more appropriate explants than the basal leaf explants. The best medium was modified MS macronutrients and micronutrients supplemented with 0.5 mg·L−1 BA and 0.1 mg·L−1 NAA (the best mean number of somatic embryos per explants was 24.10 ± 1.63). The second stage was root induction and elongation. In vitro regenerated plantlets rooted best on MS medium containing 0.1 mg·L−1 indole-3-acetic acid (IAA) and 30 g·L−1 sucrose. Rooted plantlets, following acclimatization in a greenhouse, were successfully transferred to field conditions, and 95% of the plants survived. Application of this protocol has the ability for mass multiplication, in a limited time, of the endangered species C. longgangensis.

Auxin pulse treatment holds the potential to enhance efficiency and practicability of somatic embryogenesis in potato

The objective of the current study was to simplify existing somatic embryogenesis systems in potato (Solanum tuberosum L.) cv. Desiree. The project targeted the agar-based induction phase of the potato somatic embryogenesis process as the key area for improvement. Experiments were established to ascertain the effect of a 2,4-D (2,4 dichlorophenoxyacetic acid) pulse, applied to the primary internodal section explant source and its subsequent effect on embryo induction. Parameters tested were the duration of the auxin pulse in a range from 0 to 300 min, and the concentrations of 2,4-D applied, in a range from 0 to 5,120 microM. The mean number of somatic embryos formed per explant was recorded after 4 and 8 weeks culture. Our findings indicated that the somatic embryogenesis in potato internodal segments could be evoked by an auxin (2,4-D) pulse treatment over a wide concentration and duration range. The results further suggested that a simple 20 microM 2,4-D pulse treatment could replace a lengthy 2 week induction phase in potato somatic embryogenesis and thus improve the system's practicability for wider uptake.

Induction of Somatic Embryogenesis and Plant Regeneration in Select Georgia and Pee Dee Cotton Lines

The current standard strategy for cotton transformation uses Agrobacterium for gene transfer and regeneration via somatic embryogenesis, but it is successful only for a handful of cultivars. This study was undertaken with the prospect of expanding the number of elite Upland cotton (Gossypium hirsutum L.) genotypes that can be regenerated via somatic embryogenesis. We tested 15 elite Upland cotton lines from Southeast germplasm: eight lines developed by the Georgia Agriculture Experiment Station and seven by the USDA-ARS Pee Dee cotton breeding program. These genotypes were tested on three embryo initiation-maturation media that were previously found to be capable of inducing somatic embryogenesis in diverse cotton species. Three Pee Dee lines, PD 97019, PD 97021, and PD 97100, and one Georgia line, GA 98033, responded to at least one of the three medium treatments. As expected, the regeneration efficiency of the Georgia and Pee Dee lines was relatively low as compared with the standard Coker 312 cultivar and a high degree of seed-to-seed variability was observed. However, the mean number of somatic embryos (SEs) per gram of tissues was high for the two best embryogenic lines, PD 97019 and GA 98033. Furthermore, the percentage of SEs that converted into plantlets for GA 98033 was comparable to Coker 312 for two of the three media tested.

Influence of Media Components and pH on Somatic Embryo Induction in Three Genotypes of Soybean

The influence of media components on the initiation of somatic embryogenesis in three genotypes of soybean was investigated. The following genotypes were used: Iroquois, Macon, and Savoy. Media modifications included sucrose concentration, type and concentration of auxin at two pH levels, and pH level independently. Immature cotyledons were used as the source of explant. Cotyledons were placed on a medium containing MS salts, B5 vitamins, sucrose, and auxin. Gelrite (0.2%) was used as the solidifying agent. Sucrose concentrations of 1, 2, 3, 4.5, or 6% were used. The auxins used included 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthaleneacetic acid (NAA) with each at concentrations of 45.2, 90.4, 135.7, 180.9, and 226.2 μM. The pH of each the media was adjusted to either 5.7 or 7.0 with 1 N NaOH. In an additional experiment, the effect of the two pH levels, 5.7 and 7.0, was investigated independently. Overall, the frequency of somatic embryogenesis significantly varied among the different genotypes used in this study, with Iroquois showing the highest response. Frequency of somatic embryogenesis also varied in response to the different treatments used, including sucrose and auxin. The highest initiation (91.7%) and mean number of somatic embryos per responding explant (14.9) of Iroquois was observed in a medium containing 2% sucrose. The highest initiation (97.1%) and mean number of somatic embryos per responding explant (19.5) was observed in Iroquois on 135.7 μM 2,4-D and Savoy on 135.7 μM 2,4-D, respectively, for the auxin by pH level experiment. No significant differences were observed among the two pH treatments used.

Hemoglobin Promotes Somatic Embryogenesis in Peanut Cultures

Critical parameters influencing somatic embryogenesis include growth regulators and oxygen supply. Consequently, the present investigation has focused on optimization of a somatic embryogenic system for peanut (Arachis hypogaea L.) through media supplementation with the auxin, picloram. The latter at 30 mg L(-1) was optimal for inducing regeneration of somatic embryos from cultured explants of zygotic embryos. In contrast, somatic embryogenesis did not occur in the absence of this growth regulator. An assessment has also been made of the beneficial effect on somatic embryogenesis and plant regeneration of the commercial hemoglobin (Hb) solution, Erythrogen. Hemoglobin at 1:50 and 1:100 (v:v) stimulated increases in mean fresh weight (up to a maximum of 57% over control), mean number of explants producing somatic embryos (15%) and mean number of somatic embryos per explant (29%).

Salt tolerance improvement in some wheat cultivars after application of in vitro selection pressure

Somatic embryogenesis through callus initiation has been quantified under salt stress conditions for 8 wheat cultivars currently cultivated in Morocco. The cultivars were classed according to the mean number of somatic embryos formed per immature embryo half and regenerated plants per 100 explants under saline conditions. Regenerated plants from control callus (R0–0) and callus initiated on 10 g l−1 NaCl (R0–10) did not show significant differences concerning plant height, spike length and grain number per ear but, the R0 plants remained less developed than parent plants. When watered with a solution containing more than 20 g l−1 NaCl, the seeds of cultivar Te derived from R0–10 regenerated plants exhibited the best elongation of roots and coleoptiles. Furthermore, a chlorophyll fluorescence test showed a clear improvement in salt tolerance of R0–10 plants at four to five-leaf stage, compared to R0–0 plants. It is concluded that plant regeneration from callus initiated on high NaCl levels may be a valid method of selection for salt tolerance.

Influence of growth regulators on somatic embryogenesis, plantlet regeneration, and post-transplant survival of Echinochloa frumentacea

After placement on Murashige and Skoog's basal medium supplemented with 3-5 mg/l 2,4-D, immature inflorescence expiants of Echinochloa frumentacea gave rise to three distinct types of callus: a) loosely arranged and soft; b) compact and translucent; c) compact, sticky and mucilaginous. Somatic embryo formation occurred in type 'b' callus in about 18-24 d. Callus types 'a' and 'c' did not produce somatic embryos. The highest percentage of cultures exhibiting somatic embryogenesis occurred on the medium containing 5 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos also formed directly on the inflorescence (without intervening callus formation) in about 15% of the expiants placed on this medium. The addition of paclobutrazol or uniconazole (0.25 or 1 mg/l) to the medium had no influence on the percentage of cultures exhibiting direct somatic embryogenesis, but paclobutrazol slightly increased the mean number of somatic embryos per culture. Many of the callus-derived somatic embryos germinated when subcultured on basal MS medium supplemented with kinetin. Addition of paclobutrazol or uniconazole to the culture medium at 0.25 or 1 mg/l decreased somatic embryo germination and shoot elongation but increased root length and leaf width. Both paclobutrazol and uniconazole increased survival of the plantlets following transplanting to soil. Increased post-transplant survival was accompanied by reduced water loss from plantlets produced on culture media containing triazoles.